Hydroporator: a hydrodynamic cell membrane perforator for high-throughput vector-free nanomaterial intracellular delivery and DNA origami biostability evaluation

Kizer, Megan E.; Deng, Yanxiang; Kang, Geoumyoung; Mikael, Paiyz E.; Wang, Xing; Chung, Aram J.

doi:10.1039/c9lc00041k

Cited by 55 publications

(103 citation statements)

References 39 publications

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“…S1, S2). This was different from intracellular delivery based on the mechanical deformation of the cell membrane caused by cells passing through a long, narrow microchannel [30][31][32][33] . BEAS-2B cells (human bronchial epithelium) were used to test the vacuum-assisted cell trapping.…”

Section: Vacuum-assisted Cell Patterningmentioning

confidence: 96%

On-chip multiplexed single-cell patterning and controllable intracellular delivery

et al. 2020

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Conventional electroporation approaches show limitations in the delivery of macromolecules in vitro and in vivo. These limitations include low efficiency, noticeable cell damage and nonuniform delivery of cells. Here, we present a simple 3D electroporation platform that enables massively parallel single-cell manipulation and the intracellular delivery of macromolecules and small molecules. A pyramid pit micropore array chip was fabricated based on a silicon wet-etching method. A controllable vacuum system was adopted to trap a single cell on each micropore. Using this chip, safe single-cell electroporation was performed at low voltage. Cargoes of various sizes ranging from oligonucleotides (molecular beacons, 22 bp) to plasmid DNA (CRISPR-Cas9 expression vectors, >9 kb) were delivered into targeted cells with a significantly higher transfection efficiency than that of multiple benchmark methods (e.g., commercial electroporation devices and Lipofectamine). The delivered dose of the chemotherapeutic drug could be controlled by adjusting the applied voltage. By using CRISPR-Cas9 transfection with this system, the p62 gene and CXCR7 gene were knocked out in tumor cells, which effectively inhibited their cellular activity. Overall, this vacuumassisted micropore array platform provides a simple, efficient, high-throughput intracellular delivery method that may facilitate on-chip cell manipulation, intracellular investigation and cancer therapy.

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Section: Vacuum-assisted Cell Patterningmentioning

confidence: 96%

On-chip multiplexed single-cell patterning and controllable intracellular delivery

et al. 2020

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“…This permeabilization can have lasting physiological effects, particularly in nerve cells . However, numerous human cell types, including epithelial cells, chondrocytes, and leukocytes, have demonstrated the ability to recover from mechanical compression without significant impact on viability and function …”

Section: Resultsmentioning

confidence: 99%

Cell Mechanical and Physiological Behavior in the Regime of Rapid Mechanical Compressions that Lead to Cell Volume Change

Liu

Young

et al. 2019

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Cells respond to mechanical forces by deforming in accordance with viscoelastic solid behavior. Studies of microscale cell deformation observed by high speed video microscopy have elucidated a new cell behavior in which sufficiently rapid mechanical compression of cells can lead to transient cell volume loss and then recovery. This work has discovered that the resulting volume exchange between the cell interior and the surrounding fluid can be utilized for efficient, convective delivery of large macromolecules (2000 kDa) to the cell interior. However, many fundamental questions remain about this cell behavior, including the range of deformation time scales that result in cell volume loss and the physiological effects experienced by the cell. In this study, a relationship is established between cell viscoelastic properties and the inertial forces imposed on the cell that serves as a predictor of cell volume loss across human cell types. It is determined that cells maintain nuclear envelope integrity and demonstrate low protein loss after the volume exchange process. These results define a highly controlled cell volume exchange mechanism for intracellular delivery of large macromolecules that maintains cell viability and function for invaluable downstream research and clinical applications.

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“…This method has the highest delivery efficiency and cell viability of all microfluidic techniques. [104,105] Delivery efficiency in hydroporators is a balance between delivery and viability that can be controlled through the Reynolds number of the fluid flow. Notably, nanomaterials with a variety of sizes ranging up to 2000 kDa have been delivered using a platform based on spiral vortex and vortex breakdown due to the flow at the cross and T-junctions.…”

Section: Flow-through Microfluidic Methodsmentioning

confidence: 99%

Section: Throughput and Controlmentioning

confidence: 99%

High Throughput and Highly Controllable Methods for In Vitro Intracellular Delivery

Brooks

Minnick

Mukherjee

et al. 2020

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Hydroporator: a hydrodynamic cell membrane perforator for high-throughput vector-free nanomaterial intracellular delivery and DNA origami biostability evaluation

Abstract: We present a hydrodynamic cell deformation-induced intracellular delivery platform, termed “hydroporator”.

Cited by 55 publications

References 39 publications

On-chip multiplexed single-cell patterning and controllable intracellular delivery

On-chip multiplexed single-cell patterning and controllable intracellular delivery

Cell Mechanical and Physiological Behavior in the Regime of Rapid Mechanical Compressions that Lead to Cell Volume Change

High Throughput and Highly Controllable Methods for In Vitro Intracellular Delivery

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